Simple and efficient regeneration of silica spin columns for plasmid DNA purification
Huong Thi Nguyen, Thanh Thi Duong, Hieu Van Ha, Dung Tien Nguyen, Vu Xuan Nguyen, Thuc Tri Bui
Abstract: Silica column-based DNA purification kits are widely utilized for the isolation and purification of plasmid DNA from
bacterial cultures. These kits utilize plasmid purification columns that contain a silica-based matrix, which efficiently binds DNA and enables various downstream applications such as PCR, sequencing, and cloning. While this method offers numerous advantages, it is crucial to consider the associated limitations, including the amount of DNA that can be purified and the cost of purification kits. To address these limitations, our study proposes a new method for reusing silica columns in DNA plasmid purification, based on simple washing of the used columns with ion-free water. The results demonstrated the effectiveness of washing the silica columns with water in cleaning the leftovers of plasmid DNA, resulting in DNA concentrations comparable to the negative control. Importantly, the DNA obtained from the regenerated silica columns remained suitable for downstream applications such as PCR reactions, restriction enzyme digestion, and DNA sequencing, exhibiting no significant differences compared to the newly purchased silica columns. These findings support the feasibility of reusing silica columns for DNA extraction, contributing to sustainable and cost-effective laboratory practices. By adopting this approach, researchers can maximize the utilization of silica columns while reducing the overall cost and environmental impact associated with DNA purification processes. This method provides an opportunity to optimize resources and promote eco-friendly practices in DNA research and analysis.
bacterial cultures. These kits utilize plasmid purification columns that contain a silica-based matrix, which efficiently binds DNA and enables various downstream applications such as PCR, sequencing, and cloning. While this method offers numerous advantages, it is crucial to consider the associated limitations, including the amount of DNA that can be purified and the cost of purification kits. To address these limitations, our study proposes a new method for reusing silica columns in DNA plasmid purification, based on simple washing of the used columns with ion-free water. The results demonstrated the effectiveness of washing the silica columns with water in cleaning the leftovers of plasmid DNA, resulting in DNA concentrations comparable to the negative control. Importantly, the DNA obtained from the regenerated silica columns remained suitable for downstream applications such as PCR reactions, restriction enzyme digestion, and DNA sequencing, exhibiting no significant differences compared to the newly purchased silica columns. These findings support the feasibility of reusing silica columns for DNA extraction, contributing to sustainable and cost-effective laboratory practices. By adopting this approach, researchers can maximize the utilization of silica columns while reducing the overall cost and environmental impact associated with DNA purification processes. This method provides an opportunity to optimize resources and promote eco-friendly practices in DNA research and analysis.
Keywords: efficient plasmid purification; plasmid purification; reusing silica columns; silica column-based DNA purification; sustainable laboratory practices
Citation: Nguyen, H. T., Duong, Th. T., Ha, H. V., Nguyen, D. T., Nguyen, V. X. & Bui, Th. T. (2024). Simple and efficient regeneration of silica spin columns for plasmid DNA purification. Bulg. J. Agric. Sci., 30(5), 893–898
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Date published: 2024-10-24
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